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Permanent link (DOI): https://doi.org/10.7939/R3JM7P

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A functional approach to profiling candidate genes in non model Brassicales Open Access

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Other title
Subject/Keyword
Brassicales
virus induced gene silencing
evolutionary development
heteroarthrocarpy
Brassiceae
floral symmetry
VIGS
Cleomaceae
monosymmetry
Cleome
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Mankowski, Peter J.
Supervisor and department
Hall, Jocelyn (Biological Sciences)
Examining committee member and department
Deyholos, Michael (Biological Sciences)
Cooke, Janice (Biological Sciences)
Ozga, Jocelyn (Agricultural, Food & Nutritional Science)
Department
Department of Biological Sciences
Specialization
Systematics and Evolution
Date accepted
2013-01-31T10:03:54Z
Graduation date
2013-06
Degree
Master of Science
Degree level
Master's
Abstract
Obtaining functional data is an essential component in understanding mechanisms underlying morphological variation. Virus-induced gene silencing (VIGS) is a reverse genetic technique for identifying the function of target loci through viral mediated transcript knockdown. Here we introduce this technique to two plant systems Cleome violacea (Cleomaceae) and Erucaria erucarioides (Brassicaceae) chosen for their morphological traits and phylogenetic position. In both species, a construct with a heterologous sequence and a construct with an endogenous sequence of the visual marker PHYTOENE DESATURASE (PDS) were incorporated into separate viral constructs for downregulation. Downregulation using a heterologous sequence produced a comparable fold change in transcript abundance to the endogenous sequence in both species however C. violacea was more susceptible to this process. Additional targets FRUITFULL (FUL) and a TCP1 homologue were targeted for knockdown within C. violacea to extend the application of this technique. A protocol for VIGS application is now available to both species for optimization and future functional analysis.
Language
English
DOI
doi:10.7939/R3JM7P
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
Citation for previous publication
Hall, J.C., Tisdale, T.E., Donohue, K., and Kramer, E.M. (2006). Developmental basis of an anatomical novelty: heteroarthrocarpy in Cakile lanceolata and Erucaria erucarioides (Brassicaceae). International Journal of Plant Sciences 167, 771-789.Avino, M., Kramer, E.M., Donohue, K., Hammel, A.J., and Hall, J.C. (2012). Understanding the basis of a novel fruit type in Brassicaceae: conservation and deviation in expression patterns of six genes. EvoDevo 3, 20.

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